LOCUS       sequence-181374-        6412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV DREADD expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6412)
  AUTHORS   Chai H, Diaz-Castro B, Shigetomi E, Monte E, Octeau JC, Yu X, Cohn 
            W, Rajendran PS, Vondriska TM, Whitelegge JP, Coppola G, Khakh BS
  TITLE     Neural Circuit-Specialized Astrocytes: Transcriptomic, Proteomic, 
            Morphological, and Functional Evidence.
  JOURNAL   Neuron. 2017 Aug 2;95(3):531-549.e9. doi: 
            10.1016/j.neuron.2017.06.029. Epub 2017 Jul 14.
  PUBMED    28712653
REFERENCE   2  (bases 1 to 6412)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.neuron.2017.06.029"; journalName: "Neuron"; date: 
            "2017-08-2- 2"; volume: "95"; issue: "3"; pages: "531-549.e9"
COMMENT     Sequence Label: pZac2.1 GfaABC1D rM3D mCherry SV40
FEATURES             Location/Qualifiers
     source          1..6412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    96..117
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(132..137,138..155,156..162)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    170..186
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     175..197
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     194..210
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     194..210
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     intron          1206..1338
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1321..1340
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1383..1401
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2670..2679
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2676..3386
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2821..2839)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3089..3108
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3426..3547
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3437..3456)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3491..3510
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3709..3838
                     /label=AAV2 ITR
     primer_bind     complement(3857..3874)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3857..3873)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3866..3888)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4015..4470
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4102..4121)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4312..4333
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4496..4600
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4601..4669,4670..5461)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5632..6220
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6121..6140
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
       61 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
      121 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
      181 taacaatttc acacaggaaa cagctatgac catgattacg ccagatttaa ttaaggcctt
      241 aattaggctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcggcctca
      301 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttccttgtag
      361 ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa gatctaacat
      421 atcctggtgt ggagtagggg acgctgctct gacagaggct cgggggcctg agctggctct
      481 gtgagctggg gaggaggcag acagccaggc cttgtctgca agcagacctg gcagcattgg
      541 gctggccgcc ccccagggcc tcctcttcat gcccagtgaa tgactcacct tggcacagac
      601 acaatgttcg gggtgggcac agtgcctgct tcccgccgca ccccagcccc cctcaaatgc
      661 cttccgagaa gcccattgag cagggggctt gcattgcacc ccagcctgac agcctggcat
      721 cttgggataa aagcagcaca gccccctagg ggctgccctt gctgtgtggc gccaccggcg
      781 gtggagaaca aggctctatt cagcctgtgc ccaggaaagg ggatcagggg atgcccaggc
      841 atggacagtg ggtggcaggg ggggagagga gggctgtctg cttcccagaa gtccaaggac
      901 acaaatgggt gaggggagag ctctccccat agctgggctg cggcccaacc ccaccccctc
      961 aggctatgcc agggggtgtt gccaggggca cccgggcatc gccagtctag cccactcctt
     1021 cataaagccc tcgcatccca ggagcgagca gagccagagc aggttggaga ggagacgcat
     1081 cacctccgct gctcgcaagc tttattgcgg tagtttatca cagttaaatt gctaacgcag
     1141 tcagtgcttc tgacacaaca gtctcgaact taagctgcag aagttggtcg tgaggcactg
     1201 ggcaggtaag tatcaaggtt acaagacagg tttaaggaga ccaatagaaa ctgggcttgt
     1261 cgagacagag aagactcttg cgtttctgat aggcacctat tggtcttact gacatccact
     1321 ttgcctttct ctccacaggt gtccactccc agttcaatta cagctcttaa ggctagagta
     1381 cttaatacga ctcactatag gctagcctcg aggccaccat gaccttgcac agtaacagta
     1441 caacctcgcc tttgtttccc aacatcagct cttcctgggt gcacagtccc tcggaggcag
     1501 ggctgccctt ggggacagtc actcagttgg gcagctacaa catttcacaa gaaactggga
     1561 atttctcctc aaacgacacc tccagcgacc ctctcggggg tcacaccatc tggcaagtgg
     1621 tcttcattgc cttcttaact ggcttcctgg cattggtgac catcattggc aacatccttg
     1681 tcattgtggc cttcaaggtc aacaaacagc tgaagacagt caacaactac ttcctcttaa
     1741 gcctggcctg tgcagacctg atcatcgggg tcatttccat gaacctgttc actacctaca
     1801 tcattatgaa ccgttgggca ctggggaact tagcctgcga cctctggctc tccattgact
     1861 gtgtggccag caatgcctct gtcatgaatc tgctggtcat cagctttgac aggtactttt
     1921 ccatcacttc tccattccgc taccagagcc tgatgaccag ggctcgagct ggtgtgatga
     1981 ttggtctggc ttgggtcatc tcctttgtcc tatgggctcc tgccatcttg ttctggcaat
     2041 actttgtagg gaagagaact gtgcccccag gagaatgttt cattcagttt ctgagtgagc
     2101 ccaccatcac cttcggcacg gcgatcgctg gcttttacat gcctgtcacc atcatgacta
     2161 ttttatactg gcgggtgtac cgggaggcca aggagcagat caggaagatc gaccgctgcg
     2221 agggccggtt ctatggcagc caggagcagc cgcagccacc cccgctcccc caacaccagc
     2281 ccatcctcgg caacggccgt gccagcaaga ggaagacgtc ccgtgtcatg gccatgaggg
     2341 aacacaaagc tctgcagacg ctcagtgcca tcttgctggc cttcatcatc acgtggaccc
     2401 cctacaacat catggtcctg gtgaacacct tctgtgacag ctgcataccc aaaacctatt
     2461 ggaatctggg ctactggctg tgctatatca acagcaccgt gaaccctgtg tgctatgccc
     2521 tgtgcaacaa aacattcaga accaccttca agacgctcct cttgtgccag tgtgacaaaa
     2581 ggaagaggcg caaacagcag taccagcaga gacagtcggt catttttcac aagcgagtgc
     2641 cggagcaggc cttggcggat ccaccggtcg ccaccatggt gagcaagggc gaggaggata
     2701 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     2761 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     2821 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     2881 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
     2941 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     3001 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     3061 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     3121 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
     3181 ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca
     3241 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc
     3301 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     3361 gcggcatgga cgagctgtac aagtaatcta gagtcgaccc gggcggccgc ttcgagcaga
     3421 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3481 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3541 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     3601 ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcttcctag
     3661 agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag
     3721 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     3781 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagcc
     3841 ttaattaacc taattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     3901 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     3961 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     4021 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4081 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4141 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4201 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4261 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4321 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4381 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4441 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     4501 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4561 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4621 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4681 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4741 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4801 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4861 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4921 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4981 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5041 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5101 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     5161 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5221 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5281 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5341 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5401 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5461 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5521 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5581 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5641 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5701 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5761 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5821 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5881 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5941 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6001 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6061 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6121 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6181 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6241 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6301 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     6361 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa ta
//
